Heat stabilization

{{about|a biochemistry technique|wine-making process|Winemaking#Cold and heat stabilization}}

Heat stabilization is an additive-free preservation technology for tissue samples which stops degradation and changes immediately and permanently. Heat stabilization uses rapid conductive heating, under controlled pressure, to generate a fast, homogeneous and irreversible thermal denaturation of proteins, resulting in a complete and permanent elimination of all enzymatic activity that would otherwise cause further biological changes to the tissue sample ex vivo. Due to the permanent inactivation of enzymes, heat stabilization overcomes the drawbacks of conventional tissue sample preservation techniques, such as snap-freezing followed by inhibitors.{{cite journal|date=February 2009|title=Heat Stabilization of the Tissue Proteome: A New Technology for Improved Proteomics|journal=J. Proteome Res.|volume=8|issue=2|pages=974–981|doi=10.1021/pr8006446|pmid=19159280|vauthors=Svensson M, et al.|citeseerx=10.1.1.464.2789}}

Understanding the role of proteins, peptides and small molecules in normal and diseased tissue is crucial to defining their potential use as drugs, drug targets or disease biomarkers. Yet biological changes begin the moment tissue is removed from its native environment. Dramatic alterations at the molecular level occur within seconds e.g. changed metabolism, catabolic fragmentation of large molecules (such as ATP) occurs in order to release energy, leading to disrupted control mechanisms, phosphorylation states are altered and proteins begin to degrade. As a consequence vital information may be lost or distorted, leading to inter-sample variation, risk of incorrect data interpretation and potentially misleading conclusions.{{cite journal|date=June 2013|title=The impact of biosampling procedures on molecular data interpretation|journal=Mol. Cell. Proteomics|type=Review|volume=12|issue=6|pages=1489–1501|doi=10.1074/mcp.R112.024869|pmid=23382104|vauthors=Sköld K, Alm H, Scholz B|doi-access=free |pmc=3675808}}

Heat stabilization offers significant advantages over conventional approaches to preventing biological change.{{cite journal|date=15 January 2013|title=Eliminating biological change after excision|url=http://www.genengnews.com/gen-articles/eliminating-biological-change-after-excision/4687/|journal=Gen. Eng. Biotechnol. News|type=Tutorial|volume=33|issue=2|vauthors=Söderquist M}} It can be used to replace snap freezing followed by inhibitors, pH changes, organic solvents or cross-linking. It can also be used with frozen tissue, allowing stabilization of stored samples. Heat stabilization can be used for almost any kind of tissue sample, and has been verified to be compatible with many downstream analytical techniques such as mass spectrometry,{{cite journal|date=August 2011|title=Thermal stabilization of tissues and the preservation of protein phosphorylation states for two-dimensional gel electrophoresis|journal=Electrophoresis|volume=32|issue=16|pages=2206–2215|doi=10.1002/elps.201100170|pmid=21792998|vauthors=Smejkal GB, et al.}} phospho-shotgun,{{cite journal|date=June 2012|title=Quantitative maps of protein phosphorylation sites across 14 different rat organs and tissues|journal=Nat. Commun.|volume=3|issue=876|pages=876|doi=10.1038/ncomms1871|pmid=22673903|vauthors=Lundby A, et al.|pmc=3621391|bibcode=2012NatCo...3..876L}} MALDI imaging,{{cite journal|date=July 2013|title=Localisation of adenine nucleotides in heat-stabilised mouse brains using ion mobility enabled MALDI Imaging|journal=Int. J. Mass Spectrom.|volume=345–347|pages=19–27|doi=10.1016/j.ijms.2013.02.004|pmid=|vauthors=Blatherwick EQ, et al.|bibcode=2013IJMSp.345...19B}} Western blot,{{cite journal|date=January 2013|title=Expression of trisomic proteins in Down syndrome model systems|journal=Gene|volume=512|issue=2|pages=219–225|doi=10.1016/j.gene.2012.10.051|pmid=23103828|vauthors=Spellman C, et al.}} 1D and 2D gels, reversed-phased protein arrays,{{cite journal|date=May 2013|title=Protein profiles in Tc1 mice implicate novel pathway perturbations in the Down syndrome brain|journal=Hum. Mol. Genet.|volume=22|issue=9|pages=1709–1724|doi=10.1093/hmg/ddt017|pmid=23349361|vauthors=Ahmed MM, et al.|pmc=3613160}} RIA and ELISA. The method also allows samples collected and handled in bio safety level laboratories to be subsequently handled outside such labs after treatment.{{cite journal|author=|date=August 2016|title=CDC Invests in Denator's Heat Stabilization Technology |department=News: Products & Services|journal=Gen. Eng. Biotechnol. News|volume=36|issue=14|page=8}}